Transplantation of a sheet of human corneal endothelial cell in a rabbit model
Transplantation of a sheet of human corneal endothelial cell in a rabbit model
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发表时间：
2008-01
期刊：
影响因子：
2.2
通讯作者：
Kouichiro Hitani;Seiichi Yokoo;N. Honda;T. Usui;S. Yamagami;S. Amano
中科院分区：
文献类型：
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作者：
Kouichiro Hitani;Seiichi Yokoo;N. Honda;T. Usui;S. Yamagami;S. Amano
目的建立一种新的人角膜内皮细胞（HCECs）片层的构建方法，并对HCECs片层的特性进行研究。方法将HCECs置于细胞培养板上培养1周，从细胞培养板底部加入乙二胺四乙酸以减弱HCECs的贴壁。通过用刮刀钝性地分离细胞片来构建HCEC片。将HCEC细胞片置于离体兔角膜钮扣的后表面，并移植到供体兔的角膜床上。在HCEC片组的两只眼中，用PKH 26标记培养的HCEC以观察移植后HCEC的定位。结果培养的HCECs可从培养物底部钝性脱落。ZO-1、Na+，K+-ATP酶、层粘连蛋白、纤连蛋白和IV型胶原免疫组化染色呈阳性。HCEC片材中的平均细胞密度为2，425个细胞/mm 2。HCEC片移植后，HCEC组的角膜水肿比对照组早得多。在HCEC片材组中，贴附于移植的兔角膜的后表面的单层连续细胞和移植的角膜的后表面覆盖有PKH 26标记的细胞。术后7天，HCEC片材组的平均内皮细胞密度为2，244个细胞/mm 2。结论该技术可用于角膜再生医学和角膜内皮重建。
Purpose To develop a novel method for constructing a sheet of human corneal endothelial cells (HCECs) and examine the properties of the HCEC sheet. Methods HCECs were cultured on a cell culture insert for a week; ethylenediamine tetraacetic acid was applied from the bottom of the cell culture insert to attenuate the attachment of HCECs. The sheet of HCECs was constructed by bluntly detaching the cell sheet with a spatula. HCEC cell sheets were placed on the posterior surface of excised rabbit corneal buttons and transplanted onto the corneal beds of donor rabbits. In two eyes from the HCEC sheet group, cultured HCECs were labeled with PKH26 to observe the localization of HCECs after transplantation. Results Cultured HCECs could be bluntly detached en bloc from the bottom of a culture insert. Immunostaining for ZO-1, Na+, K+-ATPase, laminin, fibronectin, and type IV collagen was positive in the cell sheet. The average cell density in a HCEC sheet was 2,425 cells/mm2. After HCEC sheet transplantation, corneal edema decreased much earlier in the HCEC group than in the control group. In the HCEC sheet group, the monolayer of continuous cells attached to the posterior surface of the transplanted rabbit cornea and the posterior surface of transplanted cornea was covered with PKH26-labeled cells. The average endothelial cell density in the HCEC sheet group seven days postoperatively was 2,244 cells/mm2. Conclusions This technique for producing an HCEC sheet might be useful in regenerative medicine for the cornea and reconstruction of the corneal endothelium.